Analyte evaluating device, and method for evaluating analyte

• US 20050069932A1
• Filed: 09/15/2004
• Published: 03/31/2005
• Est. Priority Date: 09/25/2003
• Status: Abandoned Application

• Current Assignee: Dynamic Biosensors GmbH
• Original Assignee: Fujitsu Limited
• Inventors: Tornow, Marc, Fujihara, Tsuyoshi, Arinaga, Kenji, Fujita, Shozo, Rant, Ulrich, Abstreiter, Gerhard
• Application Number:
  US10/940,746
• Publication Number:
  US 20050069932A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: G01N 2021/6441 with two or more labels
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates

Abstract

In an analyte evaluating device comprising a carrier body that can be bound with an analyte having a fluorescence-labeled part that can emit fluorescence by light received when the distance between the fluorescence-labeled part and the carrier body is enlarged, at least one factor selected from the group consisting of a light irradiation angle, a light irradiation intensity, a light irradiation area, a fluorescence detection angle, a fluorescence detection area, the shape of the carrier body, the surface area of the carrier body, a salt concentration in a medium for use in the detection, and the adhesion density of analytes on the carrier body, is made to be adjustable. A high sensitivity is realized. Evaluation is possible without introducing fluorescence-labeled parts. Evaluation for a tiny amount of sample is possible. It is also possible to evaluate objects in a mixed state. Miniaturized, complex, and integrated devices are possible.

Claims

1. An analyte evaluating device comprising:

a carrier body that can be bound with an analyte having a fluorescence-labeled part that can emit fluorescence by light received when the distance between the fluorescence-labeled part and the carrier body is enlarged, the distance between the fluorescence-labeled part and the carrier body being variable by an external action;

a light irradiation device for the fluorescence-labeled part to emit light; and

a fluorescence detecting device for detecting the fluorescence emitted by the fluorescence-labeled part,

wherein at least one factor selected from the group consisting of a light irradiation angle, a light irradiation intensity, a light irradiation area, a fluorescence detection angle, a fluorescence detection area, the shape of the carrier body, the surface area of the carrier body, a salt concentration in a medium for use in the detection, and the adhesion density of analytes on the carrier body, is adjustable.

2. An analyte evaluating device according to claim 1 wherein the distance between said fluorescence-labeled part and said carrier body can be varied by a responding part equipped on at least one of the analyte and the carrier body.

3. An analyte evaluating device according to claim 1 wherein said external action is an electromagnetic or chemical action.

4. An analyte evaluating device according to claim 3 wherein said carrier body is an electrode and said electromagnetic action is realized by applying an electric potential difference between said electrode and a counter electrode.

5. An analyte evaluating device according to claim 1 wherein said carrier body can be chemically bound with the analyte.

6. An analyte evaluating device according to claim 1 wherein said carrier body has a Au layer on the surface.

7. An analyte evaluating device according to claim 6 wherein said carrier body has an analyte binding part bound with the Au layer via a thiol group.

8. An analyte evaluating device according to claim 1 wherein said analyte has an evaluation object binding part that has a property to specifically bind to at least one evaluation object selected from the group consisting of proteins, DNAs, RNAs, antibodies, natural or artificial single-stranded nucleotides, natural or artificial double-stranded nucleotides, aptamers, products obtained by limited decomposition of antibodies with a protease, organic compounds having affinity to proteins, biomacromolecules having affinity to proteins, complex materials thereof, and arbitrary combinations thereof.

9. An analyte evaluating device according to claim 8 wherein said evaluation object is a protein.

10. An analyte evaluating device according to claim 1 wherein said responding part can be charged positively or negatively.

11. An analyte evaluating device according to claim 2 wherein said responding part comprises at least one material selected from the group consisting of proteins, DNAs, RNAs, antibodies, natural or artificial single-stranded nucleotides, natural or artificial double-stranded nucleotides, aptamers, products obtained by limited decomposition of antibodies with a protease, organic compounds having affinity to proteins, biomacromolecules having affinity to proteins, complex materials thereof, positively or negatively charged ionic polymers, and arbitrary combinations thereof.

12. An analyte evaluating device according to claim 11 wherein said responding part comprises a natural or artificial single-stranded nucleotide, or a natural or artificial double-stranded nucleotide.

13. An analyte evaluating device according to claim 11 wherein said responding part comprises a Fab fragment or (Fab)₂ fragment of an antibody.

14. An analyte evaluating device according to claim 11 wherein said responding part comprises a fragment derived from an IgG antibody, or a fragment derived from a Fab fragment or (Fab)₂ fragment of an IgG antibody.

15. An analyte evaluating device according to claim 11 wherein said responding part comprises an aptamer.

16. An analyte evaluating device according to claim 1 wherein:

said light irradiation area is not less than the surface area of the carrier body; and

said fluorescence detection area can be in the range of 80 to 120% of the surface area of the carrier body.

17. An analyte evaluating device according to claim 1 wherein:

said light irradiation area is not less than the surface area of the carrier body; and

said fluorescence detection area can be not more than half of the surface area of the carrier body.

18. An analyte evaluating device according to claim 1 wherein said carrier body is bound with said analyte.

19. An analyte evaluating device according to claim 1 wherein a plurality of the same type or different types of carrier bodies are installed.

20. An analyte evaluating device according to claim 1 wherein a plurality of the same type or different types of analytes are installed.

21. An analyte evaluating device according to claim 4 wherein:

said carrier body is an electrode; and

said responding part is a wiry formation having a diameter of not more than 100 nm that can be positively or negatively charged, is expandable and shrinkable, and is fixed on the electrode.

22. An analyte evaluating device according to claim 21 wherein at least one factor selected from the group consisting of an applied direct-current voltage, an applied alternate-current voltage, a frequency of the applied voltage, an electric potential at the center of the AC voltage, the length of the wiry formation, and the rate of adhesion of wiry formations on the carrier body is adjustable.

23. An analyte evaluating device according to claim 21 wherein said wiry formation is fixed onto the electrode by physical or chemical adsorption.

24. A method for evaluating an analyte using an analyte evaluating device according to claim 1 wherein:

the analyte is made to be bound with the carrier body;

the distance between the fluorescence-labeled part and the carrier body is varied by an external action;

a light is emitted from the light irradiation device;

the fluorescence emitted by the fluorescence-labeled part is detected by the fluorescence detecting device; and

at least one factor selected from the group consisting of a light irradiation angle, a light irradiation intensity, a light irradiation area, a fluorescence detection angle, a fluorescence detection area, the shape of the carrier body, the surface area of the carrier body, a salt concentration in a medium for use in the detection, and the adhesion density of analytes on the carrier body, is adjusted.

25. A method for evaluating an analyte according to claim 24 wherein at least one of the ratio of the light irradiation area to the surface area of the carrier body, and the ratio of the fluorescence detection area to the surface area of the carrier body, is adjusted.

26. A method for evaluating an analyte according to claim 24 wherein:

said light irradiation area is made to be not less than the surface area of the carrier body, and

said fluorescence detection area is made to be in the range of 80 to 120% of the surface area of the carrier body.

27. A method for evaluating an analyte according to claim 24 wherein:

said light irradiation area is made to be not less than the surface area of the carrier body; and

said fluorescence detection area is made to be not more than half of the surface area of the carrier body.

28. A method for evaluating an analyte according to claim 24 wherein said salt concentration in the medium is adjusted to not more than 1 M.

29. A method for evaluating an analyte according to claim 24 wherein said salt concentration in the medium is adjusted to not more than 100 mM.

30. A method for evaluating an analyte according to claim 24 wherein said analyte is made to be bound with the evaluation object before the analyte is made to be bound with the carrier body.

31. A method for evaluating an analyte according to claim 24 wherein an electrode is used as the carrier body, and an electromagnetic action is realized by applying, between said electrode and a counter electrode, an electric potential difference having a value selected from the group consisting of a constant value, a pulse value, a stepwisely changing value and a periodically changing value, or a combination thereof.

32. A method for evaluating an analyte according to claim 24 wherein at least one physical property selected from the group consisting of presence or absence of fluorescence emission, the rate of increase in the fluorescence intensity, the rate of decrease in the fluorescence intensity, the peak fluorescence intensity, the rate of change of the peak fluorescence intensity, frequency properties of the rate of change in the fluorescence intensity and a cut-off frequency of the rate of change in the fluorescence intensity is measured.

33. A method for evaluating an analyte according to claim 24 wherein the evaluation is performed, using the relationship between at least one physical property selected from the group consisting of the rate of increase in the fluorescence intensity, the rate of decrease in the fluorescence intensity, the peak fluorescence intensity, the rate of change of the peak fluorescence intensity, frequency properties of the rate of change in the fluorescence intensity and a cut-off frequency of the rate of change in the fluorescence intensity, and at least one property selected from the group consisting of a light irradiation angle, a light irradiation intensity, a light irradiation area, a fluorescence detection angle, a fluorescence detection area, the shape of the carrier body, the surface area of the carrier body, a salt concentration in a medium for use in the detection, the adhesion density of analytes on the carrier body and an applied electric potential difference.

34. A method for evaluating an analyte according to claim 24 wherein:

said carrier body is an electrode; and

said responding part is a wiry formation having a diameter of not more than 100 nm that can be positively or negatively charged, is expandable and shrinkable, and is fixed on the electrode.

35. A method for evaluating an analyte according to claim 34 wherein said analyte is evaluated by making the wiry formation come near or move away from the carrier body by applying a sine-wave or rectangular wave alternate-current electric field.

36. A method for evaluating an analyte according to claim 34 wherein at least one factor selected from the group consisting of an electric charge of the wiry formation or a substance adhered to the wiry formation, an electrostatic capacity thereof, an applied direct-current voltage, an applied alternate-current voltage, a frequency of the applied voltage, an electric potential at the center of the AC voltage, the length of the wiry formation, and the rate of adhesion of wiry formations on the carrier body is adjusted.

37. A method for evaluating an analyte according to claim 36 wherein said electric potential at the center of the AC voltage is adjusted for the carrier body electrode to have a zero voltage.

38. A method for evaluating an analyte according to claim 36 wherein said electric potential at the center of the AC voltage is changed in a stepwise manner or continuously.

39. A method for evaluating an analyte according to claim 38 wherein said electric potential at the center of the AC voltage is changed in the range between a low electric potential region where a stronger fluorescence signal is obtained, and a high electric potential region where the fluorescence signal is rapidly weakened, both of which appear when the electric potential at the center of the AC voltage is changed in a stepwise manner or continuously.